Permanently-stable, dry bacteria preparations and process of making same.



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'EMIL HELBIG, 0F FRANKFORT-ON-THE-MAIN, GERMANY, ASSIGN'OR TOIARBWERKE VORM. MEISTER LUGIUS & BR'U'NING, 0F HbCHST-ON-THE-MAIN, GERMANY, A COR- PORATION OF GERMANY.

PERMANENTLY-STABLE, DRY BACTERIA PREPARATIONS AND PROCESS OF MAKING Patented Dec. 241, 11918,

SAME.

11,288,582. Specification of Letters Patent. 1% Drawing. Application filed June 28, 1915. Serial 110.36,?42.

, in most cases, be very strongly diluted. 1t

is difiicult and troublesome for pharmacists and physicians to prepare such dilutions of an exactly determined potency, which also causes in most cases considerable waste of valuable substance. However ready-made solutions or suspensions prepared for stor* age show in practice the disadvantage that their efficiency very much diminishes after being kept in store for some time.

For instance Kochs old-tubercu1in rctains in the originial concentration its specific efliciency for many 'years without undergoing any change, whereas a dilution thereof made in the proportion of for instance 1:1000 becomes already a few days after its pro aration considerably weaker in its specific e ciency.

Attempts were made, particularly with tubercle-bacilli preparations to convert them by chemical or physical manipulations into dry preparations, that is to say into a form, for instance into tabletsif required with the addition of common salt or another injectable substance-in which even minute quantities of said preparations are stableand can easily be dosed. However the hitherto proposed working-conditions proved to be detrimental to the specific substances and the dry preparations thus obtained were soluble in water only with great difiiculty.

Now, as is known, unstable, liquid or moist substances can be brought to dryness and rendered stable, by mixing them with anhydrids of certain salts. Thus there were used by Stiikkel, Frankel and Elfer sodium sulfate and di-sodium phosphate respectively for drying serum-and other organic preparations, because it is not possible to extract in another way the water from them without considerable difliculty (see Pharmaze'utische Zentmlhalle, page 1085, statement from Wiener K Zim'sche Wochenschrift 1910 No. 43; Biochemische Zez'tschrift, Vol. 28, 1910, page 330 77.; and vol. 40, 1912, page 138 ff.) In all thecases described in the above references it does not matter at all whether or not the dry preparation obtained, which is chiefly intended to be used as an intermediate product for other purposes, is greatly increased in volume and quantity as compared with the parent-material.

However in preparing a dry tuberculin 01' other bacteria-preparation for subcutaneous injections one must endeavor to obtain the same as highly-concentrated as possible and to use only a minimum of salt, so that it may not become necessary to use later on too large quantities of the dissolving liquid and of the dilutions ready for use.

There is no ditliculty in concentrating the various tubercle-bacilli preparations in a vacuum over a dehydrating agent to such an extent that they constitute then merely a mixture of chemical and specific substances in glycerin. All attem ts hitherto made to dry and bind the said practicably suitable form without detrimentally affecting their efliciency', were in every way unsuccessful, and the solution of this problem constitutes the subject matter of my present invention.

I have ascertained that it is not possible to preparea dry tuberculin of 100 per cent.

strength neither by means of sodium phosphate nor by meansof sodium sulfate. As to sodium carbonate, although this possesses the property of drying tuberculin of high concentration, it causes a total destruction of the specific substances contained in the bacteria preparations. Guinea pigs artificially infected with tuberculosis survived even after they had been injected with from 0.1.0.15 grams of tuberculin prepared with sodium carbonate, whereas they died, like the control animals, in the course of 1-2 days after bein injected with the same quantity of liqui old tuberculin.

However it is easily, possible by means of sodium tetraborate (borax) to obtain dry preparations containing 100 per cent. and

preparations in a means of borax kills a tuberculous guineapig in the same quantity and time as does a corresponding fresh dilution of tuberculin without the addition of the said salt. The dry preparation obtained by using borax also produces typical precipitates with specific tuberculosis serum in exactly the same way as does a freshly-prepared pure tuberculin-solution of the same concentration.

The amount of specific substances contained in the tuberculin mixture may be calculated from the degree of dilution in which precipitation still occurs.

The glycerin contained in the tubercle-bacilli preparations does not constitute a pre servative'but a substituent which is a priori an indispensable condition in obtaining for instance tuberculin. It is an inconvenient admixture which greatly impedes the drying process, but which cannot be avoided, because tubercle bacilli onlygrow in a broth containing glycerin. The manufacture of a dry preparation made from a culture containing no glycerin would be by far easier and more simple, however it is entirely impracticable to withdraw the glycerin from the original tuberculinbecause this would be detrimental to its original properties and because glycerin has the effect of greatly im-' proving the solubility of the borax in water.

The glycerin added to the tubercle-bacilli emulsion is also not intended to act as a preserving, but as an emulsifying agent. Nor

does the borax" added to the tuberculin act as a preservative, but as a waterand lycerin-binding agent. There is no neef for a preservation because undiluted tuberculin is as such indefinitely stable.

The following example illustrates my in vention:

200 cc. of tuberculin containlng 20 per cent. of glycerin are concentrated in the presence of a dehydrating agent in va'cuo without heating, taking recaution to keep it sterile. After being re uced to 100 grams,

when being used, are dissolved in water in determined quantities easily to be injected.

Having now described my invention, what I claim is:

1. The process of converting bacteria preparations into permanently stable dry preparations, suitable for making injection liquids ready for use, which process consists in mixing these preparations with dehydrated borax in sufficient proportion to yield a dry preparation.

2. The process of converting bacteria preparations into permanently stable, dry preparations suitable for making injectionliquids ready for use, which process consists in mixing these preparations with dehydrated borax in presence of glycerin, the borax in sufiicient proportion to yield a dry preparation.

3. As new compositions of matter, permanently stable, dry bacteria reparations suitable for making injectioniquids ready for use, said compositions of matter consisting of a mixture of bacteria preparations with dehydrated borax.

4. As new'compositions of matter, permanently stable, dry bacteria preparations suitable for making injection-liquids ready for use, said compositions of matter consisting of a mixture of bacteria prepartions with dehydrated borax and glycerin.

5. As a new composition of matter, a permanently stable, dry tuberculin preparation suitable for making injection-liquids ready for use, said composition of matter consisting of a mixture of tuberculin with dehydrated borax, the latter in sufficient proportion to yield a dry preparation.

In testimony whereof I afiix my signature in presence of two witnesses.

DR. EMIL HELBIG.

Witnesses JEAN GR ND, CARL GRUND. 

